Systematic assessment of variability in the proteome of iPSC derivatives


The use of induced pluripotent stem cells (iPSC) to model human complex diseases is gaining popularity as it allows investigation of human cells that are otherwise sparsely available. However, due to its laborious and cost intensive nature, iPSC research is often plagued by limited sample size and putative large variability between clones, decreasing statistical power for detecting experimental effects. Here, we investigate the source and magnitude of variability in the proteome of parallel differentiated astrocytes using mass spectrometry. We compare three possible sources of variability: inter-donor variability, inter- and intra-clonal variability, at different stages of maturation. We show that the interclonal variability is significantly smaller than the inter-donor variability, and that including more donors has a much larger influence on statistical power than adding more clones per donor. Our results provide insight into the sources of variability at protein level between iPSC samples derived in parallel and will aid in optimizing iPSC studies.


Stephanie D Beekhuis-Hoekstra, Kyoko Watanabe, Josefin Werme, Christiaan A de Leeuw, Iryna Paliukhovich, Ka Wan Li, Frank Koopmans, August B Smit, Danielle Posthuma, Vivi M Heine